Objective To verify the presence of tumor necrosis factor receptor (TNF R 2) in spermatozoa of fertile men,and to provide experimental evidence for the direct action of TNF on spermatoza. Methods Using Western blotting the expressions of TNF R 2 in spermatozoa of 15 fertile men were assessed. Results TNF R 2 was expressed in spermatozoa of all the 15 fertile men. Conclusions The spermatozoa of fertile men have TNF R 2,which may transmit the cytokines signal into cytoplasm and modulate the spermatozoa function.

Objective To investigate the inhibitive effect of antisenes oligodeoxynucleotide(ASON) on the phosphodiesterase type 5(PDE5) in smooth muscle cells of human corpus cavernosum,and provide experimental groundwork for the gene therapy of erectile dysfunction. Methods PDE5 gene ASON (containing exon 1) was transfected into the corpus cavernosum smooth muscle cells with the presence of liposome DOTAP.Another sense oligodeoxynucleotide(SON) and 1% of bovine serum were also transducted into the cells as controls.PDE5 was probed and measured by Western blot at 1,2,4,6,10,24 and 48 h after transfection. Results After transfection(1～6 h), PDE5 in human corpus cavernosum smooth muscle cells was significantly lower than it was in controls( P = 0.000 ～0.014). Conclusions The results indicate that PDE5 gene antisense oligodeoxynucleotide has inhibitive effect on PDE5 in smooth muscle cells of human corpus cavernosum in vitro,and the present study provides experimental groundwork for the gene therapy of erectile dysfunction.

Objective To provide an ideal material for the investigation of penile erection. Methods Human corpus cavernosum smooth muscle cells were cultured with DMEM ( 20% of it was human serum)in vitro and the bred cells were identified by immunohistochemistry. Results After 10～14 days, spindle cells overspreaded in 6 culture bottles and they were validated as human corpus cavernosum smooth muscle cells immunohistochemically. Conclusions Human corpus cavernosum smooth muscle cells can be cultivated in feasible medium and can also become a cell line.These bred cells can serve as a material for the study of penile erection.

ObjectiveTo investigate the levels of anti-inflammatory cytokine IL-6 and pro-inflammatory cytokine IL-8 in expressed prostatic secretion(EPS)of chronic prostatitis.To study the relationship of cytokines to the mechanism of chronic prostatitis.MethodsIL-6 and IL-8 levels were measured in 102 chronic prostatitis EPS and in 28 normal EPS by radioimmunoassay.Urine was cultured and EPS was studied according to 2-glass test.NIH-CPSI(NIH-Chronic prostatitis symptom index) was performed by every patient.ResultsIL-6 and IL-8 levels were higher in Ⅱ,ⅢA and Ⅳ prostatitis (IL-6 0.51?0.57 ng/ml,IL-8 10.75?7.96 ng/ml,n=90) than those in controls (IL-6 0.32?0.51 ng/ml,IL-8 4.56?5.68 ng/ml,n=28),P0.05.There was a correlation between WBC count and IL-8 level,r=0.529,P0.05.ConclusionsIL-6 and IL-8 are elevated in EPS of Ⅱ,ⅢA and IV prostatitis, and might be considered as mediators of inflammatory response in prostate.The results indicate that IL-6 and IL-8 can be the objective parameters in the diagnosis of chronic prostatitis.

Objective To investigate the effects of mirror therapy supplemented with motor imaging training on the upper extremity functions of hemiplegic stroke patients and on their ability in the activities of daily living (ADL).Methods Forty-four stroke patients were divided into a control group and an experimental group with 22 patients in each.All were given routine rehabilitation treatment.In addition,the experimental group was given 30 minutes of mirror therapy daily supplemented with 15 minutes of motor imaging training.The control group was given routine occupational therapy.All the patients were assessed with Brunnstrom staging,the functional test for the hemiplegic upper extremity-Hong Kong (FTHUE-HK) and the Barthel index (BI) scale before therapy and after 6 weeks.Results After 6 weeks of treatment,both groups showed significant improvements in upper limb function,hand function,and ADL ability.There were significant intra-group differences and significant differences between the experimental and control groups in terms of Brunnstrom staging,FTHUE-HK scores and BI scores.Conclusions Mirror therapy supplemented with motor imaging training can benefit patients suffering hemiplegia in improving their upper limb functioning and their ADL ability.

IFIT family genes are a kind of interferon stimulated genes (ISGs), and play important roles in antiviral sector and immunity regulation. To study the regulatory effect of IFIT family genes during influenza A virus (IAV) infection, we used RNA-sequencing analysis (RNA-Seq) technique and found that when 293T cells were infected by A/WSN/33 (WSN), the concentration of IFIT family genes were increased. Further study reveals that overexpression of IFIT2 or IFIT3 could inhibit IAV replication and transcription, and cause the dose-dependent inhibition of polymerase activity of vRNP. In addition, IFIT2 and IFIT3 encoding protein could colocalize with NS1 in 293T cells infected by WSN, indicating that they might interact with each other. The results suggest that IFIT family genes can inhibit the replication and transcription of IAV, which contributes to our understanding of the regulatory effect of host factors during influenza virus infection.
HEK293 Cells ; Humans ; Influenza A virus ; physiology ; Influenza, Human ; genetics ; Intracellular Signaling Peptides and Proteins ; genetics ; Proteins ; genetics ; Virus Replication

Objective To investigate the inhibitive effect of siRNA(small interfering RNA,siRNA) on the phosphodiesterase type 5(PDE5) in smooth muscle cells of human corpus cavernosum,and provide experimental groundwork for the gene therapy of erectile dysfunction (ED). Methods Small interfering RNAs targeting PDE5 gene were systhesized by using web design software provided by Ambion,there siRNAs and control siRNA were systhesized by Ambion. SiRNAs were transfected into smooth muscle cells of human corpus cavernosum by using siPORTTM Lipid reagent;down-regulation of PDE5 mRNA was detected by RT-PCR;the inhibitive effect of PDE5 was detected by Western Blotting. Results The results of RT-PCR indicated siRNA1、siRNA2 and siRNA3 made down-regulations of PDE5 mRNA expression in the transfected groups 58.2%、14.9% and 11.8%;the PDE5 expression decreased 70.5%、19.8% and 17.3%;however the expression did not have different in control siRNA and frank group. Conclusions The synthesized siRNAs in vitro were able to down-regulate the expression of PDE5.There were different capabilities of the specific siRNAs down-regulation.It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for ED gene therapy.

Objective To study relationships between myocardial injury and the levels of serum complement C3, C4 and C5b-9 in patients with acute coronary syndrome (ACS). Methods A retrospectively analysis was conducted. 170 ACS patients [including 110 cases of ST-segment elevation myocardial infarction (STEMI) and 60 cases of non-ST-segment elevation acute coronary syndrome (NSTE-ACS)] with ischemic chest pain or chest discomfort onset within the prior 12 hours admitted to the cardiology department of Tianjin Union Medicine Center from January 2014 to July 2016 were enrolled. Thirty-six healthy cases were enrolled as control during the same time. The levels of serum complement C3, C4 and C5b-9 on 1, 3 and 7 days after admission and myocardial function indicators were analyzed. Major adverse cardiovascular events (MACE) and readmission rate were analyzed after 1 year follow-up. The correlation between serum complement levels and myocardial function indicators was analyzed by Pearson correlation analysis. Results ① The levels of serum C3, C4 and C5b-9 on the first day in NSTE-ACS group and STEMI group were significantly higher than control group [C3 (g/L): 1.04±0.33, 1.26±0.35 vs. 0.39±0.21, C4 (g/L): 0.31±0.14, 0.33±0.10 vs. 0.19±0.07, C5b-9 (g/L): 575.46±197.26, 659.26±160.77 vs. 501.40±141.51, all P < 0.05]. There were no changes of serum C3, C4 in NSTE-ACS group, but C5b-9 decreased after a peak (g/L: 700.63±218.42) at 3 days. Serum complements in STEMI group reached peak on the third day [C3 (g/L): 1.37±0.33, C4 (g/L): 0.42±0.12, C5b-9 (g/L): 754.72±136.22]. The levels of serum C4 and C5b-9 in STEMI group were higher than NSTE-ACS group on the third and seventh day. ② The levels of troponin T (TnT), creatine kinase-MB (CK-MB), solution intercellular adhesion molecule-1 (sICAM-1), global registry of acute coronary events (GRACE) scores and percutaneous coronary intervention (PCI) numbers in STEMI group were significantly higher than those in the NSTE-ACS group, which were as opposite as left ventricular ejection fraction (LVEF). However, there were no significant differences in levels of serum N-terminal pro-brain nitric peptide (NT-proBNP), Fibrinogen (Fib), readmission rate and incidence of MACE between STEMI and NSTE-ACS groups. ③ According to GRACE, patients with ACS were divided into low risk group (≤ 108 scores, 26 cases), intermediate risk group (109-140 scores, 61 cases) and highest group (> 140 scores, 83 cases). TnT and sICAM-1 in intermediate risk group were significantly increased as compared with low risk group. Levels of TnT, sICAM-1, C3, C4 and C5b-9 in the highest group were significantly higher than the low and intermediate risk groups, however the lowest LVEF was found in the highest group. ④ It was shown by Pearson correlation analyses that levels of serum C3, C4, C5b-9 were positively correlated with TnT (r value was 0.481, 0.367, 0.292, respectively, all P <0.01), sICAM-1 (r value was 0.298, 0.249, 0.365, respectively, all P < 0.01), but negatively correlated with LVEF (r value was -0.384, -0.260, -0.200, respectively, all P < 0.01). In addition sICAM-1 positively correlated with TnT (r = 0.536, P = 0.000), but negatively correlated with LVEF (r = -0.341, P = 0.001). Conclusions Serum complements activation was found in the acute phase of ACS patients. Serum complement C3, C4 and C5b-9 are involved in the process of myocardial injury, and may reflect severity of myocardial injury and cardiac dysfunction.

Objective To compare the effects of repetitive transcranial magnetic stimulation (rTMS) at various low frequencies on upper limb function after cerebral infarction.Methods Fifty patients were randomly assigned to a control group (10 cases),a sham rTMS group (10 cases) or an rTMS group which had three sub-groups treated at 0.25 Hz,0.5 Hz and 0.75 Hz with 10 cases in each.All of the patients were treated with conventional medical treatment and rehabilitation training.The sham and true rTMS groups received rTMS applied over the M1 area of the unaffected hemisphere,5 days per week for 4 weeks.Motor evoked potential (MEP) cortical latency,and central motor conduction time (CMCT) were measured and the Fugl-Meyer assessment (FMA),motricity index (MI) and a Hong Kong functional test for the hemiplegic upper extremity (FTHUE-HK) were evaluated beforehand and at Post 1 after 2 weeks of treatment and Post 2 after 4 weeks of treatment.Results The average CMCT and FMA scores of the control and sham rTMS groups both had improved significantly at Post 2.There was no significant difference in any of the indices between those 2 groups at any time point.At Post 1,the average MEP cortical latencies of the 0.25 Hz and 0.5 Hz subgroups had improved to be significantly better than those of the control and sham rTMS groups.The average CMCTs of the 0.25 Hz and 0.5 Hz rTMS subgroups were significantly shorter after treatment,and significantly better than those of the control and sham rTMS groups.At Post 2,the average MEP cortical latency of all groups except the control group showed significant improvement compared with pre-treatment.The 2 indices of the 0.25 Hz and 0.5 Hz subgroups were again significantly shorter than those of the control and sham rTMS groups,and the average CMCTs were significantly better than that of 0.75 Hz subgroup.At Post 1 the average FMA and MI scores of the rTMS subgroups had all improved significantly.In the 0.25 Hz and 0.5 Hz subgroups the average MI scores were significantly higher than those of the control and sham rTMS groups.The FTHUE-HK scores of those 2 subgroups had also improved significantly.At Post 2,the average FMA and MI scores of all groups and the FTHUE-HK scores of rTMS group had improved significantly.In the 0.25 Hz and 0.5 Hz subgroups,all of the indices were significantly better than in the control and sham rTMS groups.The average FTHUE-HK score of the 0.25 Hz subgroup was significantly superior to that of the 0.75 Hz subgroup.In the 0.75 Hz subgroup the average MI score was significantly higher than in the control and sham rTMS groups.Conclusions rTMS at either 0.25 Hz or 0.5 Hz applied to the unaffected hemisphere provides effective treatment for enhancing the excitability of the motor cortex and the motor function of a paretic upper limb after stroke.Compared with others,the total number of stimulus pulse in 0.25 Hz subgroup was the least,and priority consideration should be given to the frequency of 0.25 Hz when using rTMS in clinical treatment of cerebral infarction.

To identify adulterants from medicinal plants of Bletilla H. G. Reichenbach, the suitable candidate DNA barcoding of Bletilla was evaluated. In this study, the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the LFY homologous gene intron 2 and chloroplast ycfl gene were amplified and sequenced from forty-one samples. The intra-specific and inter-specific divergences of Bletilla were calculated, and the identification efficiency was assessed using Barcoding Gap, NJ tree by K2P distance and BLAST1 method. The result showed the intra-specific divergence of nrDNA ITS and ycJfl (0.022-0.106 and 0.017-0.106) were obviously higher than the inter-specific divergence (0-0.012 and 0-0.015), and four species of Bletilla were also accurately distinguished in NJ trees. Whereas, there was no Barcoding Gap on LFY homologous gene intron 2, thus it cannot effectively identify species of Bletilla. Using NJ tree of nrDNA ITS and ycfl gene, powdery medicine and the adulterants of Bletilla were successfully unidentified. In conclusion, nrDNA ITS and ycfl can be used as a potential DNA barcoding to identify the medicinal plants in Bletilla and its adulterants. There were only three basic differences on nrDNA ITS between "Jujing baiji" and Bletilla striata of Lu'an in Anhui province, and two basic differences in ycfl. Based on morphological and molecular data, "Jujing baiji" could be recognized as the species of Bletilla striata.